• 한국응용과학기술학회지
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• 제35권2호
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• pp.423-432
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• 2018

스피룰리나는 지구상의 가장 오래된 조류중 하나이며 피코시아닌, 토코페롤, 베타카로틴을 함유하고 있어 노화를 방지한다고 보고된 바 있다. 본 연구에서는 재배방법이 다른 스피룰리나 에탄올추출물의 UVB로 유도된 활성산소종(reactive oxygen species, ROS)의 저해와 항산화효과를 확인하였다. 실내배양 스피룰리나 에탄올추출물(ICAE)과 야외배양 스피룰리나 에탄올추출물(OCAE) 0.1, 0.5, $1mg/m{\ell}$의 DPPH 라디칼 소거능과 SOD 유사 활성을 측정하여 항산화효과를 확인하였다. UVB로 유도된 ROS의 저해 효과는 제브라피쉬 배아와 HaCaT 세포를 이용하여 확인하였다. ICAE와 OCAE 0, 0.01, 0.05, 0.1, 0.5, $1mg/m{\ell}$을 제브라피쉬 배아와 HaCaT 세포에 처리하고 UVB로 ROS를 유도하였고 DCFH-DA로 염색하여 세포내 ROS의 양을 검출하였다. 항산화효과를 측정한 결과 양성대조군인 ascorbic acid의 DPPH 라디칼 소거능은 73%, SOD 유사 활성은 86%로 나타났다. ICAE와 OCAE $1mg/m{\ell}$의 농도에서 각각 43, 57%의 DPPH 라디칼 소거능을 나타내었고, 20, 19%의 SOD 유사 활성을 나타내어 양성대조군인 ascorbic acid보다 낮았지만 유의적인 항산화효과가 있었다. UVB로 유도된 제브라피쉬 배아와 HaCaT 세포의 ROS가 음성대조군보다 증가하였고, ICAE와 OCAE 처리된 군은 농도 의존적으로 UVB만 조사된 양성대조군 보다 ROS가 감소하였다. 본 연구의 결과는 스피룰리나 에탄올추출물이 피부보호용 화장품 소재로 사용가치가 있음을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제18권3호
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• pp.404-409
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• 2008

The brown-rot basidiomycete Fomitopsis palustris is known to degrade crystalline cellulose (Avicel) and produce three major cellulases, exoglucanases, endoglucanases, and ${\beta}$-glucosidases. A gene encoding endoglucanase, designated as cel12, was cloned from total RNA prepared from F. palustris grown at the expense of Avicel. The gene encoding Cel12 has an open reading frame of 732 bp, encoding a putative protein of 244 amino acid residues with a putative signal peptide residing at the first 18 amino acid residues of the N-terminus of the protein. Sequence analysis of Cel12 identified three consensus regions, which are highly conserved among fungal cellulases belonging to GH family 12. However, a cellulose-binding domain was not found in Cel12, like other GH family 12 fungal cellulases. Northern blot analysis showed a dramatic increase of cel12 mRNA levels in F. palustris cells cultivated on Avicel from the early to late stages of growth and the maintenance of a high level of expression in the late stage, suggesting that Cel12 takes a significant part in endoglucanase activity throughout the growth of F. palustris. Adventitious expression of cel12 in the yeast Pichia pastoris successfully produced the recombinant protein that exhibited endoglucanase activity with carboxymethyl cellulose, but not with crystalline cellulose, suggesting that the enzyme is not a processive endoglucanase unlike two other endoglucanases previously identified in F. palustris.

• Asian Pacific Journal of Cancer Prevention
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• 제17권5호
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• pp.2499-2506
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• 2016

Background: Gastric cancer (GC) is the second leading cause of cancer-related death worldwide. Several environmental, genetic and epigenetic factors have been suggested to have a role in GC development. Epigenetic mechanisms like histone changes and promoter hyper-methylation are now being increasingly studied. Associations between methylation of many gene promoters with the risk of gastric cancer have been investigated worldwide. Such aberrant methylation may result in silencing of specific genes related to cell cycling, cell adhesion, apoptosis and DNA repair. Thus this molecular mechanism might have a key role in proliferation and migration of cancerous cells. Materials and Methods: In this review article we included studies conducted on DNA methylation and gastric cancer in Iranian populations. Using Science direct, Pubmed/PMC, Springer, Wiley online library and SciELO databases, all published data until 31 January 2016 were gathered. We also searched Science direct data base for similar investigations around the world to make a comparison between Iran and other countries. Results: By searching these databases, we found that the association between methylation of seven gene promoters and gastric cancer had been studied in Iran until 31 January 2016. These genes were p16, hLMH1, E-cadherin, CTLA4, $THR{\beta}$, mir9 and APC. Searching in science direct database also showed that 92 articles had been published around the world till January 2016. Our investigation revealed that despite the importance of GC and its high prevalence in Iran, the methylation status of only a few gene promoters has been studied so far. More studies with higher sample numbers are needed to reveal the relation of methylation status of gene promoters to gastric cancer in Iran. Conclusions: Further studies will be helpful in identifying associations of DNA methylation in candidate genes with gastric cancer risk in Iranian populations.

• Food Science and Biotechnology
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• 제16권3호
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• pp.392-397
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• 2007

We investigated whether ethanol extracts of the safflower seeds containing phenolic compounds were responsible for the bone-protecting effects. Crude ethanol extract (CEE) of the safflower seeds was fed for 4 weeks at the level of 1% in diet to female Sprague-Dawley rats that had been subjected to bilateral ovariectomy (OVX). The CEE effects (OVX+CEE) were evaluated by comparing results obtained from OVX, Sham, and OVX injected with $17{\beta}$-estradiol ($OVX+E_2$) groups. OVX resulted in a dramatic reduction in the trabecular bone mass of the proximal tibia (approximately 40% of the Sham group) and an increase in fat deposition in bone marrow. In $OVX+E_2$ group, the bone loss was completely prevented as well as marrow adiposity. In OVX+CEE group, approximately 80% of the bone mass was maintained compared with Sham group and fat deposition in the bone marrow was prevented. Meanwhile, the partially purified ethanol extract containing the phenolic compounds stimulated proliferation of the ROS 17/2.8 osteoblast-like cells in a dose-dependent manner, as potently as positive controls of $E_2$ and genistein. The present data demonstrate that the ethanol extracts of safflower seeds reduced bone loss caused by estrogen deficiency. The bone-protecting effect of safflower seeds seems to be mediated, at least partly, by the stimulating effect of the phenolic compounds on the growth of osteoblasts.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.297-297
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• 1994

It has been known that adrenal corticosteroids influence the expression of adrenomedullary catecholamine-synthetizing enzymes and also suppress the emission of axonal-like processes in cultured chromaffin cells. In the present study, it was attempted ta investigate the effect of 17${\alpha}$-estradiol on catecholamine(CA) secretion evoked by acetylcholine(ACh), DMPP, McN-A-343, excess K$\^$+/ and Bay-K-8644 from the isolated perfused rat adrenal gland. The perfusion of 17${\alpha}$-estradiol (10$\^$-6/ 10$\^$-4/M) me an adrenal vein for 20min produced relatively dose-dependent inhibition in CA secretion evoked by ACh (5.5 ${\times}$ 10$\^$-3/M), DMPP (10$\^$-4/M for 2min), McN-A-343 (10$\^$-4/M for 4min) and Bay-K-8644 (10$\^$-5/M for 4min), while did not affect the CA secretory effect of high K$\^$+/(5.6 x 10$\^$-2/M). Also, in the presence of 17${\beta}$-estradiol, CA secretion of ACh, DMPP and McN-A-343 without any effect on excess K$\^$+/-evoked CA secretion. However, in adrenal glands preloaded with 17${\alpha}$-estradiol (10$\^$-5/M) plus tamoxifen (10$\^$-5/M), which is known to be a selective antagonist of estrogen receptors (for 20min), CA secretory responses evoked by ACh, DMPP and McN-A-343 were considerably recovered as compared to that of 17${\alpha}$-estradiol only, but excess K$\^$+/-induced CA secretion was not affected.

• Journal of Microbiology and Biotechnology
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• 제29권12호
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• pp.2014-2021
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• 2019

Middle East respiratory syndrome coronavirus (MERS-CoV) belongs to the genus Betacoronavirus and causes severe morbidity and mortality in humans especially when infected patients have underlying diseases such as chronic obstructive pulmonary disease (COPD). Previously, we demonstrated that MERS-CoV-encoded ORF8b strongly inhibits MDA5- and RIG-I-mediated induction of the interferon beta (IFN-β) promoter activities. Here, we report that ORF8b seemed to regulate MDA5 or RIG-I differentially as protein levels of MDA5 were significantly down-regulated while those of RIG-I were largely unperturbed. In addition, ORF8b seemed to efficiently suppress phosphorylation of IRF3 at the residues of 386 and 396 in cells transfected with RIG-I while total endogenous levels of IRF3 remained largely unchanged. Furthermore, ORF8b was able to inhibit all forms of RIG-I; full-length, RIG-I-1-734, and RIG-I-1-228, the last of which contains only the CARD domains. Taken together, it is tempting to postulate that ORF8b may interfere with the CARD-CARD interactions between RIG-I and MAVS. Further detailed analysis is required to delineate the mechanisms of how ORF8b inhibits the MDA5/RIG-I receptor signaling pathway.

• IMMUNE NETWORK
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• 제2권4호
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• pp.208-216
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• 2002

Background: Reactive oxygen and nitrogen are produced by rheumatoid arthritis (RA) synovial tissue and can induce mutations in key genes. Normally, this process is prevented by a DNA mismatch repair (MMR) system that maintains sequence fidelity. Key members of the MMR system include MutS${\alpha}$ (comprised of hMSH2 and hMSH6), which can sense and repair single base mismatches and 8-oxoguanine, and MutS${\beta}$ (comprised of hMSH2 and hMSH3), which repairs longer insertion/deletion loops. Methods: To provide further evidence of DNA damage, we analyzed synovial tissues for microsatellite instability (MSI). MSI was examined by PCR on genomic DNA of paired synovial tissue and peripheral blood cells (PBC) of RA patients using specific primer sequences for 5 key microsatellites. Results: Surprisingly, abundant MSI was observed in RA synovium compared with osteoarthritis (OA) tissue. Western blot analysis of the same tissues for the expression of MMR proteins demonstrated decreased hMSH6 and increased hMSH3 in RA synovium. To evaluate potential mechanisms of MMR regulation in arthritis, fibroblast-like synoviocytes (FLS) were isolated from synovial tissues and incubated with the nitric oxide donor S-nitroso-N-acetylpenicillamine (SNAP). Western blot analysis demonstrated constitutive expression of hMSH2, 3 and 6 in RA and OA FLS. When FLS were cultured with SNAP, the RA synovial pattern of MMR expression was reproduced (high hMSH3, low hMSH6). Conclusion: Therefore, oxidative stress can relax the DNA MMR system in RA by suppressing hMSH6. Decreased hMSH6 can subsequently interfere with repair of single base mutations, which is the type observed in RA. We propose that oxidative stress not only creates DNA adducts that are potentially mutagenic, but also suppresses the mechanisms that limit the DNA damage.

• Asian-Australasian Journal of Animal Sciences
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• 제28권4호
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• pp.573-583
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• 2015

B-32 is one of a panel of monoclonal anti-idiotypic antibodies to growth hormone (GH) that we developed. To characterize and identify its potential role as a novel growth hormone receptor (GHR) agonist, we determined that B-32 behaved as a typical $Ab2{\beta}$ based on a series of enzyme-linked immunosorbent assay assays. The results of fluorescence-activated cell sorting, indirect immunofluorescence and competitive receptor binding assays demonstrated that B-32 specifically binds to the GHR expressed on target cells. Next, we examined the resulting signal transduction pathways triggered by this antibody in primary porcine hepatocytes. We found that B-32 can activate the GHR and Janus kinase (2)/signal transducers and activators of transcription (JAK2/STAT5) signalling pathways. The phosphorylation kinetics of JAK2/STAT5 induced by either GH or B-32 were analysed in dose-response and time course experiments. In addition, B32 could also stimulate porcine hepatocytes to secrete insulin-like growth factors-1. Our work indicates that a monoclonal anti-idiotypic antibody to GH (B-32) can serve as a GHR agonist or GH mimic and has application potential in domestic animal (pig) production.

• 생명과학회지
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• 제14권1호
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• pp.26-31
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• 2004

본 연구는 redox regulator로 알려 진 Ref-1 (Redox factor-1)과 결합하는 새로운 단백질을protein-protein interaction의 원리를 이용한 방법인 yeast two-hybrid assay로 검색, 동정하고, 검색된 단백질의 in vitro, in vivo 기능을 규명하는 데 그 목적을 두고, mouse 11-day Embryo cNA library를 prey로, full length REF-1을 bait로 하여 yeast strain 인 HF7C에 cotransformatiom시킨 후 histidine, leucine, tryptophan이 결핍된 SD plate에서 키워 자란 yeast transformants를 $\beta$-galactosidaseassay하여 screening하여 분리한 세 개의 clone중 한 clone이 DNA sequencing으로 확인한 결과 mouse thioredoxin임을 확인하였다.

• Animal cells and systems
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• 제15권3호
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• pp.189-196
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• 2011

We investigated the in vitro effects of nonylphenol (NP) and 3,3',4,4',5-pentachlorobiphenyl (PCB126) on steroidogenesis in redlip mullet, Chelon haematocheilus, oocytes. In experiment 1, we investigated the effects of NP and PCB126 on steroid production from exogenous steroid precursors. Vitellogenic oocytes (0.75 mm in diameter) were incubated with 10 and 100 ng/ml NP or PCB126 with $[^3H]17{\alpha}$-hydroxyprogesterone as a precursor. The major metabolites produced were androstenedione, testosterone (T), estrone, and estradiol-$17{\beta}$ ($E_2$). Both NP and PCB126 increased T production and decreased $E_2$ production, except for 100 ng/ml PCB126. In experiment 2, oocytes (0.65-0.75 mm in diameter) were exposed to NP and PCB126 at different concentrations (0.01, 0.1, 1, 10, and 100 ng/mL). After the incubation, T and $E_2$ production was measured by radioimmunoassay. NP inhibited $E_2$ production at concentrations of 0.01 and 0.1 ng/ml in 0.75-mm-diameter oocytes. NP at 1 and 100 ng/mL stimulated T production, but had no observable effect on $E_2$ production. PCB126 treatment did not affect $E_2$ production at any of the concentrations tested. NP alone at 0.1 ng/mL resulted in a significant decrease in $E_2$ production in 0.65-mm-diameter oocytes. PCB126 did not show any significant effects on either T or $E_2$ production at all concentrations tested. These results suggest that NP acts like an antiestrogen at lower concentrations (0.01-0.1 ng/ml) in vitellogenic oocytes of redlip mullet.